链霉菌ZG0429的分类鉴定与链霉亲和素的分离纯化研究

从土壤中筛选得到1株高产链霉亲和素的放线菌ZG0429,根据形态观察、培养特征、生理生化鉴定以及16S rRNA序列分析,初步判定该菌株为链霉菌属中的淡紫灰链霉菌(Streptomyces lavendulae)。经发酵,ZG0429的链霉亲和素产量可达201.0mg/L。进一步采用硫铵沉淀和凝胶过滤层析纯化,链霉亲和素的回收率为76.87%,纯度可以达到97.03%。该方法简单易行,成本低廉,可得到高产量、高纯度、高活性的目的蛋白,为链霉亲和素发酵产品的大规模纯化提供了依据。     

Disruption of the Fatty Acid Δ<Superscript>6</Superscript>-Desaturase Gene in the Oil-Producing Fungus <Emphasis Type="Italic">Mortierella isabellina</Emphasis> by Homologous Recombination

The γ-linolenic acid-producing fungus Mortierella isabellina 6-22 is an important industrial strain. To clarify the biosynthetic pathways for polyunsaturated fatty acids in this strain, a disruption vector pD4MI6, including 5′ and 3′ regions of the fatty acid Δ⁶-desaturase open reading frame as homologous recombination elements and the Escherichia coli hygromycin B (HmB) phosphotransferase gene (hph) as selectable marker, was successfully constructed. Following transformation of pD4MI6 into the hygromycin B-sensitive recipient strain M. isabellina 6-22-4, a Δ⁶-desaturase gene-defective mutant strain was selected that was unable to produce γ-linolenic acid as determined by gas chromatography and molecular analysis. The morphology and physiology of the mutant, such as colony shape, color, and growth rate, were changed dramatically compared with that of strain M. isabellina 6-22-4.     

一个新葡萄糖淀粉酶基因的克隆与表达

根据已报道的米根霉葡萄糖淀粉酶基因序列,通过PCR方法,从天然少根根霉的总DNA中克隆到含有四个内含子的葡萄糖淀粉酶基因。通过设计引物并采取重叠PCR方法删除内含子,获得了新的少根根霉葡萄糖淀粉酶(Rhizopus arrhizu glucoamylase,RaGA)cDNA序列(Accession number:DQ903853)。该基因在毕赤酵母中成功表达,表达产物具有较高的葡萄糖淀粉酶活性。     

以潮霉素B抗性为选择标记的深黄被孢霉原生质体转化

利用亚硝基胍(MNNG)诱变方法筛选了一株深黄被孢霉潮霉素B敏感型菌株M6-22-4。采用PEG介导的方法,将含有E.coli潮霉素B抗性标记的PD4质粒转入敏感株M6-22-4原生质体,并在潮霉素B浓度为400μg/mL的选择培养基上筛选转化子,获得了1.6~2.8个转化子/μg质粒DNA的转化频率。稳定性实验表明,质粒线性化后所获得的转化子在PDA培养基上传代10代以后,转接到选择平板上有31.6%仍具有HmB抗性;随机挑选了3个转化子,通过PCR方法检测到潮霉素抗性基因的存在,Southern杂交发现,潮霉素抗性基因已经以1~2拷贝数整合到深黄被孢霉M6-22-4染色体上,这是深黄被孢霉转化系统的首次报道。     

Synthesis, characterization and biological activity of complexes of lanthanum(III) with 2-(1'-phenyl- 2'-carboxyl-3'-aza-n-butyl)-1,10-phenanthroline and 2-(1'-p-phenol-2'-carboxyl-3'-aza-n-butyl)-1,10-phenanthroline

Two novel ligands 2-(1'-phenyl-2'-carboxyl-3'-aza-n-butyl)-1,10-phenanthroline (L1) and 2-(1'-p-phenol-2'-carboxyl-3'-aza-butyl)-1,10-phenanthroline (L2), and their La(III) complexes of La(III)L1, La(III)(L1)(2), La(III)L2, and La(III)(L2)(2), were synthesized and characterized by (1)H NMR, elemental analysis, IR, thermal analysis and conductance measurement. All complexes have been assayed for anticancer activity in vitro against HL-60 (human leukocytoma) cells, PC-3MIE8 (human prostate carcinoma) cells, BGC-823 (human stomach carcinoma) cells, MDA-MB-435 (human galactophore carcinoma) cells, Bel-7402 (human liver carcinoma) cells, and HeLa (human cervix carcinoma) cells. Results showed that the two complexes La(III)L1 and La(III)(L1)(2) exhibited good cytotoxic activity against different cell lines in general; and La(III)(L1)(2) is more effective than cisplatin against all six cell lines. DNA-binding studies indicated that, besides the intercalation, the complexes bind to DNA by the other interaction(s).     

Amperometric glucose biosensor based on self-assembly hydrophobin with high efficiency of enzyme utilization

Hydrophobins are a family of natural self-assembling proteins with high biocompability, which are apt to form strong and ordered assembly onto many kinds of surfaces. These physical-chemical and biological properties make hydrophobins suitable for surface modification and biomolecule immobilization purposes. A class II hydrophobin HFBI was used as enzyme immobilization matrix on platinum electrode to construct amperometric glucose biosensor. Permeability of HFBI self-assembling film was optimized by selecting the proper HFBI concentration for electrode modification, in order to allow H₂O₂ permeating while prevent interfering compounds accessing. HFBI self-assembly and glucose oxidase (GOx) immobilization was monitored by quartz crystal microbalance (QCM), and characterization of the modified electrode surface was obtained by scanning electron microscope (SEM). The resulting glucose biosensors showed rapid response time within 6 s, limits of detection of 0.09 mM glucose (signal-to-noise ratio = 3), wide linear range from 0.5 to 20 mM, high sensitivity of 4.214 × 10⁻³ A M⁻¹ cm⁻², also well selectivity, reproducibility and lifetime. The all-protein modified biosensor exhibited especially high efficiency of enzyme utilization, producing at most 712 μA responsive current for per unit activity of GOx. This work provided a promising new immobilization matrix with high biocompatibility and adequate electroactivity for further research in biosensing and other surface functionalizing.     

内蒙古高原荒漠区几种锦鸡儿属(Caragana)优势植物的生理生态适应特性

比较了内蒙古高原荒漠区4种锦鸡儿属(Caragana)优势植物——柠条锦鸡儿、狭叶锦鸡儿、垫状锦鸡儿和荒漠锦鸡儿的生理适应特性。研究发现:4种锦鸡儿属植物中,荒漠锦鸡儿的叶片含水量、束缚水/自由水比值、水势以及水分利用效率最低,气孔导度日变化表现为与气温变化相似的早晚低、中午高的单峰曲线。4种锦鸡儿蒸腾速率日变化状况相似,均在10:00达到最大,以后逐渐降低,但日蒸腾积累值垫状锦鸡儿<荒漠锦鸡儿<柠条锦鸡儿<狭叶锦鸡儿。相比较,柠条锦鸡儿、狭叶锦鸡儿和垫状锦鸡儿对当地的水分条件有良好的适应性,能够保持较好的水分状况,水分亏缺在15%以下,而荒漠锦鸡儿叶水分亏缺日变幅较大,保水能力不及其它3种。渗透调节比较研究发现:荒漠锦鸡儿的渗透势最低,细胞质离子浓度最高,无机渗透调节物产生的渗透势所占的比例也更大,说明其低渗透势的维持主要来自无机渗透调节物质的较多积累。保护酶和自由基比较研究发现:POD和SOD活性荒漠锦鸡儿明显高于其它3个种,但CAT活性在4种植物中无显著差别;叶片自由基含量狭叶锦鸡儿>荒漠锦鸡儿>柠条锦鸡儿>垫状锦鸡儿。这些结果表明:(1)荒漠锦鸡儿对干旱环境的适应方式与其它3种不同,柠条锦鸡儿、狭叶锦鸡儿和垫状锦鸡儿以强保水能力、维持稳定的水分而适应,而荒漠锦鸡儿以更负的渗透势、更高细胞质离子浓度弥补其更多的水分消耗和对水分变化的强耐性而适应,较高的保护酶活性可能是其强耐性的生理基础之一。(2)保水能力弱的锦鸡儿种主要通过无机离子的积累,调节细胞质渗透势,保持水分平衡,这是一种相对节省能量的适应对策。     

内蒙古中东部草原羽茅Epichlo属内生真菌的分布及rDNA-ITS序列系统发育

对内蒙古中东部地区分布的羽茅6个地理种群的染菌率进行了调查,采集种子并从中分离得到不同形态型的内生真菌,选取其中的19株进行rDNA-ITS片段的扩增、克隆、测序和系统发育分析。结果表明：（1）6个样地羽茅种群内生真菌感染率除西乌旗为96.7%外,其他5个样地均为100％,表明内生真菌侵染羽茅并非偶然现象,二者之间存在一种稳定的共生关系。（2）ITS和5.8S序列得到的N-J树显示,相对于Epichlo属的其他参考菌株,不同地理种群羽茅中的内生真菌聚为一类,形成一个具有97％支持强度的分支。由此推测,不同地理种群羽茅中的内生真菌具有相同的起源点。（3）结合形态观察结果和rDNA-ITS序列分析结果可以看出,羽茅内生真菌种群的优势种亲缘关系较近,可能起源于同一种内生真菌;但由于其地理分布广、气候差异大、群落类型差别也较大,从而造成不同地理种群内生真菌形态上的分化以及种群间明显的遗传分化和较高的遗传多样性。     

金羽蛾属一新种记述(鳞翅目,羽蛾科)

记录了采自天津的1新种郑氏金羽蛾Agdistis zhengi sp.nov. （图1 -3）。该种分布于天津,与直钩金羽蛾A.uncirectangula Hao ＆ Li（图4）相似,但可以通过以下特征与后者区别：爪形突的左侧一支端部近圆形深凹,两侧顶端尖,均向外侧弯曲;抱器背臂整体看起来象鹅的头部;阳茎基环两叶,小而不明显;囊形突大,近环带状,末端圆;第8腹板末端突起的形状和位置与后者不一致等。文中给出了新种成虫图,以及雌雄外生殖器特征图。模式标本保存在南开大学生命科学学院昆虫标本室。 新种以著名昆虫学家郑哲民教授的姓氏命名。     

中国螟蛾科一新纪录属及一新种记述(鳞翅目,螟蛾科,斑螟亚科)

首次报道桂斑螟属Gunungia在中国的分布,首次报道该属的雌性个体,并据此对属征进行了补充描述。桂斑螟属目前只包括模式种Gunungia rimba Roesler ＆ Küppers,分布于印度尼西亚苏门答腊岛。本文记述了采自我国广西、贵州和云南的1新种——凹头桂斑螟Gunungia capitirecava sp.nov.,编制了已知种的检索表,提供了新种的特征图。模式标本均保存在南开大学生命科学学院昆虫标本室。 凹头桂斑螟,新种Gunungia capitirecavasp.nov.（图1 -7） 翅展16.0 -24.5mm。新种与G.rimbaRoesler ＆ K櫣ppers近似,但新种雄性头顶被白色V型鳞毛簇;抱器背在端部1/6处具一齿状突起,阳茎基环两侧臂粗壮,末端圆钝。 正模♂,广西防城港峒中林场, 640m, 2002-04-08 ,郝淑莲、薛怀军采（玻片号RYD04343）。副模： 7 ♂♂,1♀,贵州赤水桫椤, 240m, 2000-09-21 -23 ,于海丽; 1 ♀,云南瑞丽珍稀植物园, 1 000m, 2005-08-05 ,任应党采; 1 ♂,1♀,云南西双版纳自然保护区补蚌管理站,650m,2005—08—25,任应党采。